Method for screening for pluripotent stem cell growth-promoting factor

Abstract

This invention relates to a method for screening for a growth-promoting factor for pluripotent stem cells with a conditioned medium which is generated by culturing feeder cells in a serum-free medium that contains L-ascorbic acid, insulin, transferrin, selenium, and sodium bicarbonate and does not contain a serum nor serum replacement; a method for growing pluripotent stem cells via feeder-free culture using the conditioned medium; and a method for growing pluripotent stem cells by carrying out feeder-free culture of pluripotent stem cells cultured in advance on feeder cells in the serum-free medium.

The invention claimed is: 
     
       1. A method for growing pluripotent stem cells comprising carrying out feeder-free culture of pluripotent stem cells in a conditioned medium which is generated by culturing feeder cells in a serum-free medium that contains L-ascorbic acid, insulin, transferrin, selenium, and sodium bicarbonate and does not contain a serum nor serum replacement. 
     
     
       2. The method according to  claim 1 , wherein the serum-free medium is DMEM/F12 medium containing L-ascorbic acid, insulin, transferrin, selenium, and sodium bicarbonate. 
     
     
       3. The method according to  claim 1 , wherein the culturing of the feeder cells is carried out with the addition of growth factors to the serum-free medium. 
     
     
       4. The method according to  claim 1 , wherein the culturing of the feeder cells is carried out without the addition of growth factors to the serum-free medium and the feeder-free culture of pluripotent stem cells is carried out with the addition of a growth factor to the conditioned medium. 
     
     
       5. The method according to  claim 2 , wherein the culturing of the feeder cells is carried out with the addition of growth factors to the serum-free medium. 
     
     
       6. The method according to  claim 2 , wherein the culturing of the feeder cells is carried out without the addition of growth factors to the serum-free medium and the feeder-free culture of pluripotent stem cells is carried out with the addition of a growth factor to the conditioned medium. 
     
     
       7. The method according to  claim 3 , wherein the growth factor is FGF2 and/or TGF-β1. 
     
     
       8. The method according to  claim 5 , wherein the growth factor is FGF2 and/or TGF-β1. 
     
     
       9. The method according to  claim 1 , wherein the feeder cells are mouse embryonic fibroblasts. 
     
     
       10. The method according to  claim 1 , wherein the pluripotent stem cells are ES cells or iPS cells.